FCS2 Closed Chamber Inverted

FCS2 Closed Chamber-Inverted

After rigorous preparation your cells will need a micro-observation environment that is conducive to their viability, compatible with your experiments protocol and all techniques of microscopy!

What is it?

The Focht Chamber System 2 (FCS2 ®) is a closed system, live-cell micro-observation chamber, that offers several advantages over other chambers. In addition to its unique perfusion and thermal control systems it is fully compatible with all modes of microscopy. It is also the only chamber to combine high-volume laminar flow perfusion rates with Koehler illumination and precise temperature control without an air curtain.

The system is comprised of:

  • Chamber (environmental optical cavity, this page)
  • Electronic Controller (this page)
  • Stage Adapter

Accessories recommended for immersion objectives:

How does it work?

Temperature Control
The FCS2 was designed to maintain accurate thermal control and allow high-volume laminar flow perfusion. Both of these functions are incorporated into our patented Microaqueduct Slide. The surface of the slide, opposite the specimen side, is coated with an electrically conductive transparent thin film of Indium-Tin Oxide (ITO) and two electrical contacts (busbars). When the FCS2 is completely assembled two electrical contacts, (not shown in drawing), which are contained in the electrical enclosure rest on the busbars. A temperature controller is used to pass a regulated current flow through the ITO Coating. This causes the surface of the slide to heat. The heat is transferred through the perfusable media to the cell surface on the coverslip thereby providing first surface thermal control. The self locking base of the chamber is also temperature regulated to provide peripheral heat as well.

Microaqueduct Perfusion
A fluid pathway is formed by separating the Microaqueduct slide from the coverslip containing cells with a single silicone gasket. This gasket can be any thickness from 50 micron to 1mm and any lateral geometry you choose or create. This arrangement allows the user to define the flow characteristics. Therefore, you are not limited by the geometry of the optical cavity instead you select or create it! Fluid access to this flow channel is made through two 14-gauge needle stock tubes protruding from the sides of the chamber top. These tubes provide fluid connection to two perfusion holes in the Microaqueduct slide that interface two “T” shaped grooves cut into the inner surface of the Microaqueduct slide. The “T” groove allows the media to seek the path of least resistance and become nearly laminar before flowing across the cells. This technique eliminates the need for the metal perfusion ring and additional gaskets, which are the limiting factors, required by most conventional chambers.

Microaqueduct design enables proper Koehler illumination with high-numeric aperture optics for both transmitted and reflected modes of microscopy.

  • Suitable for no flow through high rate flow procedures where a rapid exchange of media is required with low cell surface shear
  • Cell temperature can be controlled from ambient to 50 degrees C +/- 0.2 degrees C without the need of an air curtain
  • Temperature is controlled uniformly across entire field with media equilibrating as it enters the chamber
  • Closed system so that bicarbonate CO2 or organic buffers can be employed
  • Compatible with 1/16″ tubing for perfusion (C-Flex, Tygon, etc.)
  • Easily assembled with ordinary skill (no tools required)
  • Stand-alone temperature controller with an alarm circuit to safeguard your cells
  • Near laminar flow
FCS2

Isometric View of Optical Cavity

Microaqueduct Slide is transparent glass but rendered as a solid to improve contrast to illustrate function. This image is rendered upside down, to demonstrate how the Optical Cavity works. In an FCS2 chamber the coverslip will be on the bottom.

Isometric View of Optical Cavity
Isometric View of Optical Cavity

Assembled FCS2

FCS2 Exploded View

FCS2 Exploded View
FCS2 Exploded View

Singular Lower Gasket

By simply changing this one gasket you can change the volume of the chamber. This gasket can have any internal geometry you desire and can be any thickness from .1mm to 1mm. The drawing below shows the standard shapes of the gaskets that we include with every FCS2. We also include solid gaskets for you to custom fit to your application. Once you have found the shape that works best for your experiment you can contact us to have a die made to those specifications. Examples of standard gasket outlines (below)

Singular Lower Gasket

The Gasket Set Includes:

(3) 0.1mm 30mm Round
(2) 0.1mm 14 x 24
(3) 0.25mm 30 mm Round
(2) 0.25mm 14 x 24
(1) 0.25mm Blank
(3) 0.5mm 30mm Round
(2) 0.5mm 14 x 24
(2) 0.5mm Blank
(5) 0.75mm 30mm Round w/holes
(2) 0.75mm 14 x 24
(1) 0.75mm Blank
(2) -1.0mm 30mm Round
(1) 1.0mm 14 x 24
(1) 1.0mm Blank

Custom shapes are available. Simply contact Bioptechs to make arrangements for their production.

Open Mode Top for the FCS2

The open mode option allows for the FCS2 to be assembled without the microaqueduct slide thus exposing the cell on the cover slip for microinjection. The coverslip can then be removed and reassembled with the microaqueduct slide for long term, time-lapse.

Open Mode Top for the FCS2

Open Mode Adapter Installed on FCS2 Base

Open Mode Top for the FCS2

Open Mode Adapter Installed on FCS2 Base

Additional Resources

If you are using immersion objectives on mammalian specimens, you will need an Objective Heater!

Click here to see a video of FCS2 assembly and demonstration

Click here for a PDF of FCS2 user instructions

Click here for a PDF about Traditional Stage Warming with thermal images.

Click here for a PDF about Typical FCS2 Perfusion Configuration For Induced Change Biology

To assist you in selecting the most appropriate system to meet your needs, please download and fill out a PDF of Micro-Environmental System Profile Questions

Specifications

Physical Size:
75mm OD 13mm high

Coverslip No:
1.5 thick x 40mm Diameter

Imaging Aperture:
22mm

Maximum Volume:
706mm^3

Minimum Volume:
<31mm^3

Maximum Volume Exchange Rate:
1/sec

Minimum Fluid Aperture:
0.6mm^2

Separation between optical surfaces:
50 – 1000 microns

External port ID:
1.6mm

Temperature Stability:
+/- 0.2 degrees C